Identification and enzymatic evaluation of peptidase producing bacteria
Subject Areas : Molecular Microbiology
Mojtaba Mortazavi
1
,
Batool Khani Sakhvidi
2
,
Masoud Torkzadeh-Mahani
3
,
safa lotfi
4
,
محمود ملکی
5
,
مهدی رحیمی
6
1 - Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
2 - Department of Biodiversity, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
3 - Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
4 - Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
5 - Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
6 - Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
Keywords: Protease enzyme, srRNA 16 gene, phylogenetic tree, protease activity, enzyme-producing bacteria ,
Abstract :
Background & Objectives: Proteases, hydrolases used extensively in industries like detergents and textiles, are particularly valuable when sourced from microbes due to their resilience. This study aims to identify bacteria capable of producing proteases and to investigate their enzymatic properties. Understanding the function and efficiency of these enzymes can lead to process improvements and novel applications in various industries.
Materials & Methods: Bacterial protease-producing bacteria were isolated from water and soil samples collected in Kerman and screened on SMA based on halo diameter. The species exhibiting the highest protease activity was selected, its 16S rDNA sequenced, and identified using BLAST. Protease activity was then evaluated at different temperatures.
Results: Enzyme tests show that sample 11b has protease activity in the temperature range of 25-45°C. After bacterial DNA extraction, PCR processing, and epidermal sequencing, Staphylococcus vitellinus, Bacillus toyonensis, Citrobacter jillini, and Staphylococcus sevieri were identified.
Conclusion: This analysis introduces a new bacterial species that can be used in various industrial and clinical fields.
References:
1. Kumar D, Savitri TN, Verma R, Bhalla TJRJM. Microbial proteases and application as laundry detergent additive. 2008;3(12):661-72.
2. King JV, Liang WG, Scherpelz KP, Schilling AB, Meredith SC, Tang W-J. Molecular basis of substrate recognition and degradation by human presequence protease. Structure. 2014;22(7):996-1007.
3. Shen Y, Joachimiak A, Rosner MR, Tang W-J. Structures of human insulin-degrading enzyme reveal a new substrate recognition mechanism. Nature. 2006;443(7113):870-4.
4. Ibrahim ASS, Al-Salamah AA, El-Toni AM, Almaary KS, El-Tayeb MA, Elbadawi YB, et al. Enhancement of alkaline protease activity and stability via covalent immobilization onto hollow core-mesoporous shell silica nanospheres. International journal of molecular sciences. 2016;17(2):184.
5. Razzaq A, Shamsi S, Ali A, Ali Q, Sajjad M, Malik A, et al. Microbial proteases applications. 2019;7:110.
6. Shah D, Mital KJAit. The role of trypsin: chymotrypsin in tissue repair. 2018;35:31-42.
7. Garrido C, Wollman F-A, Lafontaine IJGB, Evolution. The evolutionary history of peptidases involved in the processing of organelle-targeting peptides. 2022;14(7):evac101.
8. Ahvenainen T. Ubiquitin-specific protease 14 in Huntington's disease. 2015.
9. Buller AR, Townsend CA. Intrinsic evolutionary constraints on protease structure, enzyme acylation, and the identity of the catalytic triad. Proceedings of the National Academy of Sciences. 2013;110(8):E653-E61.
10. Babic M, Jankovic P, Marchesan S, Mausa G, Kalafatovic DJJoci, modeling. Esterase sequence composition patterns for the identification of catalytic triad microenvironment motifs. 2022;62(24):6398-410.
11. Shafee T. Evolvability of a viral protease: experimental evolution of catalysis, robustness and specificity: University of Cambridge; 2014.
12. Pietzner M, Wheeler E, Carrasco-Zanini J, Cortes A, Koprulu M, Wörheide MA, et al. Mapping the proteo-genomic convergence of human diseases. 2021;374(6569):eabj1541.
13. Shamsi TN, Parveen R, Fatima SJIjobm. Characterization, biomedical and agricultural applications of protease inhibitors: A review. 2016;91:1120-33.
14. Domsalla A, Melzig MF. Occurrence and properties of proteases in plant latices. Planta medica. 2008;74(07):699-711.
15. Rao CS, Sathish T, Ravichandra P, Prakasham RSJPB. Characterization of thermo-and detergent stable serine protease from isolated Bacillus circulans and evaluation of eco-friendly applications. 2009;44(3):262-8.
16. Rani K, Rana R, Datt SJIJCLS. Review on latest overview of proteases. 2012;2(1):12-8.
17. Mohammad Mashhadi-Karim MA, Seyyed Latif Mousavi Gargari,, Sarshar M. Comparison of function of immobilized and free Bacillus licheniformis cells in production of alkaline protease. Journal of Microbial World. 2011;4(1):22-15.-- (In Persian)
18. Arastoo Badoei-dalfard PA, Narjes Ramezanipour, Zahra Karami1, Batool Ghanbari. The production of alkaline protease by Bacillus tequilensis FJSH2 isolated from Jiroft’s slaughterhouse wastes. Journal of Microbial World. 2015;8(1):54-61.-- (In Persian)
19. Matkawala F, Nighojkar S, Kumar A, Nighojkar AJWJoM, Biotechnology. Microbial alkaline serine proteases: production, properties and applications. 2021;37:1-12.
20. Hamza TA. Bacterial protease enzyme: Safe and good alternative for industrial and commercial use. Int J Chem Biomol Sci. 2017;3(1):1-10.
21. Mohammad Mashhadi-Karim MA, Seyyed Latif Mousavi Gargari, Meysam Sarshar. Comparison of function of immobilized and free Bacillus licheniformis cells in production of alkaline protease. Journal of Microbial World. 2011;4(1):15-22.
22. Mrudula SJAB, Microbiology. A Review on Microbial Alkaline Proteases: Optimization of Submerged Fermentative Production, Properties, and Industrial Applications. 2024;60(3):383-401.
23. Shanmugaiah V, Mathivanan N, Balasubramanian N, Manoharan PJAJoB. Optimization of cultural conditions for production of chitinase by Bacillus laterosporous MML2270 isolated from rice rhizosphere soil. 2008;7(15).
24. Bamdad F, Shin SH, Suh J-W, Nimalaratne C, Sunwoo HJM. Anti-inflammatory and antioxidant properties of casein hydrolysate produced using high hydrostatic pressure combined with proteolytic enzymes. 2017;22(4):609.
25. Soltanmoradi R, Razmara AS, HOSEINI DSR. Molecular screening and of extracellular protease producing Serratia marcescens from the different environments of west Mazandaran. 2013.-- (In Persian)
26. Karray F, Ben Abdallah M, Kallel N, Hamza M, Fakhfakh M, Sayadi SJMBR. Extracellular hydrolytic enzymes produced by halophilic bacteria and archaea isolated from hypersaline lake. 2018;45(5):1297-309.
27. Flandrois J-P, Perrière G, Gouy M. leBIBI QBPP: a set of databases and a webtool for automatic phylogenetic analysis of prokaryotic sequences. BMC bioinformatics. 2015;16(1):251.
28. Kumar S, Stecher G, Li M, Knyaz C, Tamura KJMb, evolution. MEGA X: molecular evolutionary genetics analysis across computing platforms. 2018;35(6):1547-9.
29. Solanki P, Putatunda C, Kumar A, Bhatia R, Walia AJB. Microbial proteases: ubiquitous enzymes with innumerable uses. 2021;11(10):428.
30. Song P, Zhang X, Wang S, Xu W, Wang F, Fu R, et al. Microbial proteases and their applications. 2023;14:1236368.
31. Sharma V, Tsai M-L, Nargotra P, Chen C-W, Kuo C-H, Sun P-P, et al. Agro-industrial food waste as a low-cost substrate for sustainable production of industrial enzymes: a critical review. 2022;12(11):1373.
32. Majithiya VR, Gohel SDJAB, Biotechnology. Agro-industrial Waste Utilization, Medium Optimization, and Immobilization of Economically Feasible Halo-Alkaline Protease Produced by Nocardiopsis dassonvillei Strain VCS-4. 2024:1-25.
33. Zarparvar P, Amoozegar MA, Fallahian MRJC, Research M. Diversity of culturable moderate halophilic and halotolerant bacteria in Incheh Boroun hyper saline wetland in Iran. 2014;27(1):44-56.- (In Persian)
34. Babavalian H, AMOUZGAR M, Pourbabaei A. Isolation, Identification and Characterization of moderately halophilic bacteria producing Hydrolytic enzymes from Aran-Bidgol salt Lake. 2009.-(In Persian)
35. Patil R, Jadhav B. Isolation and characterization of protease producing Bacillus species from soil of dairy industry. Int J Curr Microbiol App Sci. 2017;6(6):853-60.
36. Reshma R. Effect of alkaline protease produced from fish waste as substrate by Bacillus clausii on destaining of blood stained fabric. Journal of Tropical Life Science. 2021;11(1).
37. Saxena S, Verma J, Modi DR. RAPD-PCR and 16S rDNA phylogenetic analysis of alkaline protease producing bacteria isolated from soil of India: Identification and detection of genetic variability. Journal of Genetic Engineering and Biotechnology. 2014;12(1):27-35.
38. Sims G. Nitrogen starvation promotes biodegradation of N-heterocyclic compounds in soil. Soil Biology and Biochemistry. 2006;38(8):2478-80.
39. He W, Zhang M, Jin G, Sui X, Zhang T, Song FJME. Effects of nitrogen deposition on nitrogen-mineralizing enzyme activity and soil microbial community structure in a Korean pine plantation. 2021;81:410-24.
40. Rao MB, Tanksale AM, Ghatge MS, Deshpande VV. Molecular and biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev. 1998;62(3):597-635.
